To improve the quality of healthcare, it is desirable that diagnostic tests such as immunoassays are quickly and inexpensively conducted at the point of care such as at home or by the hospital bed. To that end, microfluidic devices are ideal as they require only very small quantities of samples and reagents, thereby reducing cost and space.
Fluorescence methods in microfluidic immunoassay devices currently involve bulky optical detection systems that typically focus excitation light from an external light source onto a sample in a microchannel, and collect any fluorescence emitted with a set of complex lenses, mirrors and optical filters. As fluorescence emissions are isotropic, collection efficiency is generally low, usually less than 5%. Improving efficiency usually means needing a more complex, bigger and more expensive optical system. Alignment of the excitation light with the sample and the detection system is another challenge given the narrow channels in microfluidic device. This is exacerbated when excitation and collection is to be done along the length of the channel in order to obtain the total fluorescence emitted in a channel. Isotropic fluorescence emissions and scattered excitation light may also propagate through the microfluidic substrate and produce cross-talk in adjacent channels for multi-channel devices. Fluorescence background noise from the microfluidic substrate may even be higher than emissions produced by the samples.